The present invention provides methods for generating mammalian T cell populations comprising antigen-specific CD4+CD25+ regulatory T cells from freshly isolated CD4+CD25− T cells. The method comprises selecting CD4+CD25− T cells from a sample obtained from a mammalian subject; determining the MHC Class II type of the subject; inducing the generation of antigen-specific regulatory T cells by contacting the isolated CD4+CD25− T cells in a culture vessel with an induction agent for a time period sufficient to generate antigen-specific CD4+CD25+ regulatory T cells; and selecting the CD4+CD25+ antigen-specific regulatory T cells by sorting the cells in the induction culture with a selection agent comprising at least one artificial multimeric MHC Class II/peptide complex that corresponds to the MHC Class II type of the subject.